Point-of-Care Rapid RNA test for HIV?Phase 2 CrossLife Technologies Inc. Project Summary/Abstract Current antibody point-of-care tests cannot distinguish between a mother's and baby's antibodies to detect if an infant has become infected with Human Immunodeficiency Virus (HIV). According to a recent WHO/UNAIDS/UNICEF report, only a predicted 28 percent of babies born to HIV-positive mothers obtained an HIV test within the first two months [1]. Therefore, there are unmet needs for new and renovated test that can be applied at the point-of-care to detect HIV infections. A rapid, sensitive, specific simple and cost-effective paper strip molecular test that detects HIV at the point-of-care was developed by integrating TARA chemical and device innovations. The target sequences are amplified using the TARA (Template Assisted Rapid Assay) technology which is composed of a template-dependent chemical transfer reaction with high turnover, gold nanoparticle-HRP, and lateral flow readout. The test should be rapid (<30 minutes), and results in the appearance of colored bands that are detectable by eye. The resulting cartridge similar to a pregnancy test will automate all steps of molecular analysis and amplification for detection of HIV RNAs in blood finger-stick without sample preparation. In Phase 1, we demonstrated 100% sensitivity and specificity for our assay on a small number of simulated clinical samples. This is well above levels reported in studies of the Alere Q and SAMBA point of care HIV assays [52-53]. The goal of this Phase 2 proposal is to 1) Paper microfluidics for TARA assay automation, 2) verify the performance of our TARA HIV test using a large number of clinical samples (n=400), and then 3) pursue World Health Organization (WHO) expedited review approval and recommendation for procurement on our product. After initial marketing of the test developed in Phase 2, we plan to pursue CLIA waiver in order to allow for untrained, lay persons to operate the test. This approach is largely due to the extended budget and timeline that will be required to cover further development of our test to make it CLIA-waived. Our test will be more rapid than the RT-PCR tests because the results will be available at the point-of-need and not in a laboratory where the instrument is located.